ARRANGEMENT OF MESSENGER-RNA AT THE DECODING SITE OF HUMAN RIBOSOMES - 18S-RIBOSOMAL RNA NUCLEOTIDES AND RIBOSOMAL-PROTEINS CROSS-LINKED TO OLIGOURIDYLATE DERIVATIVES WITH ALKYLATING GROUPS AT EITHER THE 3' OR THE 5' TERMINI
ARRANGEMENT OF MESSENGER-RNA AT THE DECODING SITE OF HUMAN RIBOSOMES - 18S-RIBOSOMAL RNA NUCLEOTIDES AND RIBOSOMAL-PROTEINS CROSS-LINKED TO OLIGOURIDYLATE DERIVATIVES WITH ALKYLATING GROUPS AT EITHER THE 3' OR THE 5' TERMINI
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DOI：
10.1111/j.1432-1033.1994.tb20100.x
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发表时间：
1994-12-01
期刊：
影响因子：
--
通讯作者：
KARPOVA, GG
中科院分区：
文献类型：
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作者：
MALYGIN, AA;GRAIFER, DM;KARPOVA, GG
本文研究了在同源Phe-tRNA(Phe)存在下，人胎盘80S核糖体与长达12个尿苷残基的核糖体的亲和标记，即在5‘端带有4-(N-2-chloroethyl-N-methylamino)benzylmethylphosphamide基团或在3’端带有2‘，3’-O-[4-(N-2-chloroethyl-N-methylamino)]benzylidene残基的寡尿酰基的烷基化衍生物。所有的信使核糖核酸类似物都只修改了40S亚基。经5‘-端烷基化基团类似的信使核糖核酸修饰的18S rRNA的含量随着试剂低聚尿苷含量的增加而显著降低。用逆转录技术测定了与该mRNA类似物烷基化的18S rRNA的核苷酸。对于与3‘-末端烷基化基团类似的mRNA，确定了G1702和G1763/G1764为交联点。逆转录酶终止对应的条带的强度取决于试剂寡尿酸部分的长度。对于(PU)(3)和(PU)(4)衍生物，与18S rRNA 5‘-末端烷基化基团的类似物的交联位分别为A1023、C1026、C1057和A1058，对于(PU)(6)衍生物为单核苷酸C1057。核糖体蛋白S26被发现是与(PU)(6)和(PU)(12)相同的衍生物修饰的主要目标。
Affinity labeling of human placental 80S ribosomes with mRNA analogs of up to 12 uridyl residues, i.e. alkylating derivatives of oligouridylates bearing either 4-(N-2-chloroethyl-N-methylamino)benzylmethylphosphamide group at the 5'-termini or 2',3'-O-[4-(N-2-chloroethyl-N-methylamino)]benzylidene residue attached to the 3'-termini, in the presence of cognate Phe-tRNA(Phe) has been investigated. All the mRNA analogs modified only the 40S subunit. The fraction of 18S rRNA modified by the mRNA analogs with the alkylating group at the 5'-end decreased dramatically with extension of the reagent oligouridylate moiety. Nucleotides of 18S rRNA alkylated with the mRNA analogs were determined using a reverse transcription technique. For the mRNA analogs with the alkylating groups at the 3'-termini, G1702 and G1763/G1764 were identified as the cross-linking sites. The intensities of the bands corresponding to reverse transcriptase stops depended on the length of the reagent oligouridylate moieties. Cross-linking sites of the mRNA analogs with the alkylating group at the 5'-termini on 18S rRNA were A1023, C1026, C1057 and A1058 for the (pU)(3) and (pU)(4) derivatives and a single nucleotide C1057 for the (pU)(6) one. Ribosomal protein S26 was found as the main target of modification with the same derivatives of (pU)(6) and (pU)(12).